analySIS image-analytical software allows quick determination of vascular growth stimulation.
Cardiovascular and cancer diseases are the number one and two causes of death in the developed world. The vascular biology field has gained global significance in the search for new medical treatments and effective preemptive measures. Research in this field focuses on blood vessels. Specifically, this means examining the growth of blood vessels (referred to as "angiogenesis") as well as any vascular change of a pathological nature. Anti-angiogenic tumor therapies are an example of the new therapeutical opportunities resulting from this area of research. These types of therapies exploit the fact that tumors require nutrients and oxygen supplied by blood vessels in order to be able to grow. The aim is not directly attacking the tumor itself, rather selectively attacking the blood vessels supplying it. Their growth is inhibited in order to cut the tumor off from its `supply lines.' The search for anti-angiogenic substances (ie, substances that inhibit vascular growth) is the fundamental first step toward the development of such therapies.
Searching for inhibitors systematically
In the Department of Vascular Biology & Angiogenesis Research of the Tumor Biology Center in Freiburg, Germany, a testing technique has been developed for quickly and reliably determining whether or not a substance is anti-angiogenic or pro-angiogenic (meaning that it stimulates vascular growth). This technique is based on a new kind of cell cultivation method in which endothelial cells (a layer of cells which make up the lining of vessels) are shaped into three-dimensional cell spheroids. First, they are embedded in collagen gel. Next, the substances being tested are placed in the same collagen gel. If the substance has a pro-angiogenic effect, a sprouting of capillaries can be observed under the microscope. Tiny vascular structures as fine as strands of hair grow out of the spheroids. To determine whether or not a substance is anti-angiogenic, it is blended with a second substance known to be pro-angiogenic. If the first substance (the one being tested) weakens or completely suppresses the known pro-angiogenic effect of the second substance, this first substance is evidently anti-angiogenic.
Counting and measuring
The question is, just how pro- or anti-angiogenic is the substance being tested (or, how pro- or anti-angiogenic is the entirety of all the blended substances being tested)? And another question is, to what extent does the anti-angiogenic effect depend on the pro-angiogenic substance used in the blend? In order to be able to quantify this effect, there is some serious counting and measuring to be done. How many capillary sprouts have grown? How long are they? The pro-angiogenic effect is measured according to "additive sprout length. "This refers to the total length of the vessels that have grown from a cell spheroid and is measured in micrometers. To ensure that all this measuring and statistical evaluation can be carried out as quickly as possible, the latest digital technology is used.
Using analySIS to determine "additive sprout length"
For each test, ten spheroids embedded in collagen gel are acquired under an inverse microscope (Olympus IX50) using a digital camera (Olympus DP50).The acquisitions are read directly into the PC by Soft Imaging System's analySIS image-analytical software. Then they can be quickly and precisely evaluated right at the PC. The software recognizes the microscope magnification selected and automatically calibrates the images accordingly. Then the measuring within the acquired images begins--all onscreen. This involves simply measuring the lengths of the various vascular sprouts. All measurement data is automatically entered into a sheet and statistically evaluated (including the additive sprout length).
Further evaluation and automation All images and data can be:
* archived in the database via drag and drop
* inserted into a report template of the user's design or
* exported into another application program.
Transferring statistical data into a sheet calculation program such as Microsoft Excel is not a problem. To further develop their testing technique, the researchers use additional image-processing and image-analytical functions available within the analySIS software. Using morphological filters and an automatic particle analysis they are able to automatically record the perimeter of the spheroids (including the vascular sprouts). The length of the perimeter reflects the extent of the pro-angiogenic effect. It is therefore just as suitable as the additive sprout length for measuring this effect. Due to automation, this testing technique takes even less time, making the technique well-suited for fast analysis of many substances (referred to as `semi-high throughput screening'). A professional service based on this testing technique has been developed. It is used by biotech companies for testing substances for their anti-angiogenic properties (www.spherogenex.de).
Rainer Maurer Prize for Innovation
Dr. Thomas Korff, a human biologist, has been conferred the Rainer Maurer Prize for Innovation for the development of this new digital image analysis technique by the renowned Gesellschaft fur Zell- und Gewebezuchtung e.V. (GZG), the German chapter of the European Tissue Culture Society. The jury's decision was based on the grounds that this testing technique, "represented the development of an innovative cell-culture method suitable for testing biologically-pharmacologically active substances and which is of potential significance with regard to the treatment of disease".
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Peter Stallknecht, Ph.D., is responsible for Public Relations at Soft Imaging System. Thomas Korff Ph.D., is Leader of the Department of Vascular Biology & Angiogenesis Research of the Tumor Biology Center in Freiburg, Germany. They may be contacted at editor@scimag.com.
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